Mice were euthanized 24 hours post RD, eyes were enucleated, embedded in O.C.T. compound (Tissue Tek; Sakura Finetek, Torrance, CA, USA) and fresh-frozen at −80 °C. Eyes were then cut in the sagittal plane at 20 µm-thickness on a cryostat (Leica CM1850; Leica Biosystems, Buffalo Grove, IL, USA) and serial sections were collected on polyethylene terephthalate-membrane (PET) frame slides (PET FrameSlide #0010; steel frames, RNase-free, material number #11505190, Leica Microsystems, Wetzlar, Germany). Sections were fixed in 75% ethanol (30 seconds), washed with nuclease-free water (30 seconds), stained with 0.02% toluidine blue solution for 20 seconds and washed again as described above. Finally, sections were dehydrated with 75%, 95% and 100% ethanol (30, 30 and 2 × 30 seconds respectively). LCM was performed with the Leica LMD7000 system and LMD application version 7.5 (Leica Microsystems, Wetzlar, Germany). Photoreceptors’ layer was cut by laser and collected into 0.5 ml tubes containing RNAlater stabilization solution (Invitrogen, Thermo Fisher Scientific, Waltham, MA, USA).
Lmd application version 7
Manufactured by Leica
Sourced in Germany, United States
LMD application version 7.5 is a software package designed for the control and operation of Leica's laser microdissection systems. The core function of this application is to provide a user interface for the precise targeting and extraction of specific cellular or tissue samples from microscope slides for further analysis.
Automatically generated - may contain errors
Lab products found in correlation
Lmd7000 system, by Leica (2 mentions)
Cm1850, by Leica (1 mentions)
Rnalater stabilization solution, by Thermo Fisher Scientific (1 mentions)
Tissue tek, by Sakura Finetek (1 mentions)
Toluidine blue solution, by Merck Group (1 mentions)
Nuclease free water, by Merck Group (1 mentions)
Frame slide, by Leica (1 mentions)
Nuclease free water, by Thermo Fisher Scientific (1 mentions)
Rnalater, by Thermo Fisher Scientific (1 mentions)
Oct compound, by Sakura Finetek (1 mentions)
2 protocols using lmd application version 7
1
Laser Capture Microdissection of Photoreceptors
2
Laser Capture Microdissection of Mouse Retina
Check if the same lab product or an alternative is used in the 5 most similar protocols
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LCM was performed with the Leica LMD7000 system and LMD application version 7.5 (Leica Microsystems, Buffalo Grove, IL, USA). Enucleated mouse eyes were frozen in O.C.T Compound (Sakura Finetek, Torrance, CA, USA) and cut into 20 μm sections on Frame slides (Leica Microsystems). The sections were fixed with 75% ethanol for 30 s, washed with nuclease-free water (Life Technologies) for 30 s, stained with 0.02% toluidine blue solution (Sigma-Aldrich) for 20 s, and washed with nuclease-free water for 30 s. The sections were then dehydrated with 75 and 95% ethanol for 30 s each, and twice with 100% ethanol for 30 s. The tissues from the ONL, subretina (where the sensory retina detached from the RPE), and the RPE (with choroid) were cut by laser and collected into 0.5 ml tubes containing RNAlater (Life Technologies).
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